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Localization of o ‐glycosylation sites of MUC1 tandem repeats by QTOF ESI mass spectrometry
Author(s) -
Hanisch FranzGeorg,
Green Brian N.,
Bateman Robert,
PeterKatalinic Jasna
Publication year - 1998
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/(sici)1096-9888(199804)33:4<358::aid-jms642>3.0.co;2-3
Subject(s) - chemistry , tandem mass spectrometry , glycosylation , mass spectrometry , tandem , tandem repeat , chromatography , biochemistry , materials science , genome , composite material , gene
The potential of electrospray mass spectrometry (ESMS) for the sequencing of glycopeptides was evaluated using quadrupole time‐of‐flight (QTOF) technology in the MS/MS mode. The location of O ‐glycosylation sites was possible in the positive ion (+) mode by detection of prominent y‐and b‐fragment ions from the underivatized TAP25‐2 [T 1 APPAHGVT 9 S 10 APDT 14 RPAPGS 20 T 21 APPA], an overlapping sequence of MUC1 tandem repeats which had been glycosylated in vitro by two GalNAc residues in the positions T9 and T21. The high mass resolution and accuracy of QTOF‐(+)ESMS allowed reliable structural assignments. The reduced complexity of the fragment spectra and the higher signal‐to‐noise ratio render QTOF‐(+)ESMS an alternative mass spectrometric approach to the identification of O ‐glycosylation sites when compared with sequencing by post‐source decay matrix‐assisted laser desorption/ionization MS. Diagnostic ions from the N ‐terminus in the b‐series offered direct evidence, which was supported by indirect evidence from the C ‐terminus ions of the y‐series. The higher glycosylated GalNAc 2 ‐substituted fragments were mainly observed as multiply ionized species. © 1998 John Wiley & Sons, Ltd.